Aflatoxin contamination in wheat flour samples from Golestan province, Northeast of Iran

Background: Due to the high toxicity of aflatoxin and its effects on public health, determination of aflatoxin level in Wheat flour samples in the Golestan province, north of Iran was investigated. To examine the effect of seasonal changes, summer and winter sampling was performed with standard sampling methods. Methods: A total of 200 flour samples were collected from 25 factories. HPLC method with immunoaffinity chromatography was used to measure aflatoxin types (G2, G1, B2 and B1). Statistical analysis was performed by the Pearson correlation test, One-way ANOVA and multivariate regression analysis. Results: Mean total aflatoxin levels of samples were 0.82 and 1.99 ng/g in summer and winter, respectively. Aflatoxin B1 levels were detected in 3.1%, 7.4% over permissible limits by worldwide regulations in samples collected in summer and winter, respectively. Aflatoxins in winter were higher than summer. The highest frequency of aflatoxin contamination in winter was B2 (98%) and in summer G1 (51%). The relationship between humidity and rate of aflatoxin B1 and total aflatoxin was significant in winter. Results of multivariate regression were showed the strongest relationship with humidity and aflatoxin level. Despite the contamination of flour samples, there was no contamination higher than the standard limit of Iran Standard Institute. But it was significantly higher than similar studies from other regions. Conclusions: Therefore, with regard to negative impacts of aflatoxin on health, aflatoxin contamination should be considered in future programs. Decrease of aflatoxin contamination may be made practical through reducing wheat storage duration and controlling humidity

Complex regulation of the aflatoxin biosynthesis gene cluster of Aspergillus flavus in relation to various combinations of water activity and temperature

A microarray analysis was performed to study the effect of varying combinations of water activity and temperature on the activation of aflatoxin biosynthesis genes in Aspergillus flavus grown on YES medium. Generally A. flavus showed expression of the aflatoxin biosynthetic genes at all parameter combinations tested. Certain combinations of aw and temperature, especially combinations which imposed stress on the fungus resulted in a significant reduction of the growth rate. At these conditions induction of the whole aflatoxin biosynthesis gene cluster occurred, however the produced aflatoxin B1 was low. At all other combinations (25 °C/0.95 and 0.99; 30 °C/0.95 and 0.99; 35 °C/0.95 and 0.99) a reduced basal level of cluster gene expression occurred. At these combinations a high growth rate was obtained as well as high aflatoxin production. When single genes were compared, two groups with different expression profiles in relation to water activity/temperature combinations occurred. These two groups were co-ordinately localized within the aflatoxin gene cluster. The ratio of aflR/aflJ expression was correlated with increased aflatoxin biosynthesis

Quantitative determination of aflatoxin by high performance liquid chromatography in wheat silos in Golestan province, north of Iran

Background: Aflatoxins are the most common mycotoxins that contaminate crops. They are produced by fungi such as Aspergillus flavus and Aspergillus parasiticus. Wheat (Tricitumaestivum) is one of the most important staple foods used in Iran, and the environmental conditions in the north of Iran are favorable to fungal growth. This study was designed in order to determine the aflatoxin concentration in wheat samples from silos in Golestan Province north of Iran. Methods: Samples were collected from three silos of Golestan province. First, aflatoxins were isolated using immu-noaffinity chromatography. Then the aflatoxin concentrations were determined by High performance liquid chroma-tography (HPLC) method and fluorescence detector. Results: Ten out of 34 samples (29.4 of samples) were contaminated by aflatoxins.No concentration was found above permitted aflatoxin levels in Iran (15 ng/g). In one sample (2.9), aflatoxin B1 was seen over the permissible limits in Iran. The highest level found in samples for total aflatoxin, aflatoxin B1, aflatoxin B2, aflatoxin G1 and aflatox-in G2 were 7.08 ng/g, 6.91 ng/g, 0.29 ng/g, 1.37 ng/g and 0.23 ng/g, respectively. No correlation was found between humidity levels in wheat samples contained aflatoxin and wheat samples without aflatoxin. Conclusion: Despite the total aflatoxins determined in samples were below the permissible limits in Iran, the 29 aflatoxin contamination rate can negatively affect health factors and it should not be neglected. So, it is predictable that if the storage duration of samples increases, the aflatoxin contamination levels will increase. © 2015, Iranian Journal of Public Health. All rights reserved

High-performance liquid chromatography and Enzyme-Linked Immunosorbent Assay techniques for detection and quantification of aflatoxin B1 in feed samples: a comparative study.

ObjectiveComparison was done between high-performance liquid chromatography (HPLC) and a competitive enzyme-linked immunosorbent assay (ELISA) for detection and quantification of aflatoxin B1 (AFB1) in feed samples. The two procedures were standardized and validated before the actual experiment. Five concentrations (0, 5, 10, 20 and 30 ppb) of feed samples were used for both methods. For the HPLC technique, the samples were extracted in acetonitrile/water (90/10) solution, cleaned-up using solid phase extraction (SPE) column, and derivatized by water/trifluoroacetic acid/glacial acetic acid (35/10/5) solution before instrument analysis. The samples were extracted in 70% methanol for the ELISA technique.ResultsThe two tests showed very strong linearity with correlation coefficient value of > 0.99 using standard solutions. The mean recovery rate was 92.42% (with relative standard deviation (RSD) of 5.97) and 75.64% (RSD = 34.88) for HPLC and ELISA, respectively. There was no statistically significant difference in recovery rate between the two methods. There was a positive correlation (r = 0.84) between them which indicated that the two techniques can be used to detect and quantify aflatoxin B1 in feed samples. However, there were variations among replicates for the ELISA method, which shows that this method is more applicable for screening purposes

Mycoflora and Aflatoxin Contamination of Some Foodstuffs

Analysis was made of the mycoflora and aflatoxin contamination of Rice (Oryza sativa), Beans (Phaseolus vulgaris), Corn (Zea mays), and Groundnut (Arachis hypogaea) sold in four different markets in Sango-Ota, Ogun state, Nigeria. Sixty four samples comprising of four samples of each foodstuff from four food vendors in four different markets was assayed. The samples were contaminated with different species of fungi to include Aspergillus flavus, Aspergillus tamarii, Aspergillus niger, Rhizopus nigricans, Rhizopus oryzae, Saccharomyces cerevisiae, Aspergillus parasiticus, Fusarium moniliforme, Fusarium verticilliodes, Aspergillus ochraceus, Cladosporium cladosporioide, Mucor spp, Trichodema spp, Rhizopus arrhizus and Aspergillus fumigates. Aspergillus flavus and Fusarium spp had the highest rate of occurrence among the isolated fungi. Aflatoxins B1, B2, G1 and G2 were found associated with the samples at concentration ranging from 9 - 25 ppb, 8 - 12 ppb, 6 - 21 ppb, 4 - 8 ppb respectively. The fungal counts were between 6.3 x 102 to 7.0 x 103 cfu/g. The moisture content and the pH of samples were between 10.9 to 28.0% and 6.20 to 6.66 respectively. Effective storage and adherence to HACCP principles will help prevent contamination of foodstuffs with aflatoxigenic fungi

Mycoflora of fungal contamination in wheat storage (silos) in golestan province, north of Iran

Background: Cereal products are susceptible to mould damage during pre- and post-harvesting stages of the production. The regional specificity of Golestan province in the northern region of of Iran, with its high temperature and high relative humidity, acts as a leading factor for the growth of aflatoxin-producing fungi. It is well known that contamination of starch-based ingredients with mycotoxigenic fungi is a risk factor among the consumers due to its aflatoxins. Objectives: This survey was carried out to determine the extent of fungal contamination of wheat in three silos of Golestan province in Iran. Materials and Methods: 34 samples from three active silos were collected in order to clean the polyethylene bags. Wheat analyzed for fungal contamination and aflatoxins extracted by immunoaffinity column chromatography, and measured by HPLC method. Results: The most common moulds isolated were Alternaria spp. 26.7%, Aspergillus niger 21.4%, Fusarium spp. 17.8%, Aspergillus flavus 10.7%, Cladosporium spp. 10.7%, Penicillium spp. 8.9%, and Rhizopus spp. 3.5%. The screening of aflatoxin, B1, B2, G1 and G2 was carried out. 10(29.4%) samples of wheat had traces of aflatoxin, but in a level lower than the standard levels [Institute of Standards and Industrial Research of Iran (ISIR< 15 ng/g)]. Conclusions: Despite the lower detected aflatoxin levels (lower than the ISIR level), the fungal contamination rate could not be neglected. Since the isolated mycotoxigenic fungi such as Aspergillus spp. and Fusarium spp. are important in food industry, it would be possible that the increased retention time of samples might have raised the detected contamination rate. © 2013, Ahvaz Jundishapur University of Medical Sciences

Dietary aflatoxin exposure and impaired growth in young children from Benin and Togo: cross sectional study

Fetal and early childhood environment, including the nutritional status of the pregnant mother and the infant, are considered critical for growth and risk of disease in later life. Many people in developing coun­ tries are not only malnourished but also chronically exposed to high levels of toxic fungal metabolites (mycotoxins). One family of mycotoxins, the aflatoxins, are carcinogenic and immunotoxic and cause growth retardation in animals. Aflatoxins contaminate staple foods in West Africa, particularly maize and ground­ nuts, as a result of hot, humid storage conditions that promote fungal growth. High exposure to aflatoxins occurs throughout childhood in the region, suggest­ ing that growth and development could be critically affected.We assessed exposure to aflatoxins in relation to anthropometric measures in children in Benin and Togo

Exposición crónica a hongos productores de aflatoxinas relacionada a daños hepáticos en chinchillas (chinchilla lanigera) destinadas a la producción de piel

Chinchilla pelt is a rare and expensive fur. Therefore, breeding these animals is a profitable activity. Confirmed acute cases of aflatoxin intoxication have been reported in Argentinean farms. The aims of this study were i) to evaluate mycobiota and AFB1 -producing species in chinchilla feeds ii) to investigate their natural AFB1 contamination and iii) to analyze histopathological lesions in chinchilla livers. Feed samples (A: fur chinchillas, B: mother chinchillas, C: lucerne cubes) were collected from a factory and a farm. Livers of sacrificed chinchilla from the farm were macroscopically and microscopically examined. Total fungal counts of feed C exceeded 1x104 CFU g-1. Aspergillus, Fusarium and Penicillium were the prevalent genera, while A. flavus, A. fumigatus, F. verticillioides and F. proliferatum were the prevalent species. 50 % of A. flavus strains from factory samples and 69.7 % from farm samples produced 2.78 to 8. 64 µg g-1 and 0.66 to 58.8 µg g-1 AFB1 , respectively. Aflatoxin B1 was detected only in feeds from the farm, finding the highest incidence in feed C. Toxin levels varied between 1.90 and 97.34 µg kg-1 AFB1 . Mean levels in feed A and C exceeded 20 µg kg-1. Macroscopic examination of livers revealed normal appearance, size and color. However, histopathological examination indicated 63.3 % showed slight to moderate lipid degeneration with diffuse cytoplasm vacuolation, 9 % intense lipid cytoplasm vacuolation and 27.3 % hydropic degeneration and nuclear vacuolation in hepatocytes. A periodic monitoring of aflatoxins in feeds and their ingredients can prevent acute outbreaks and economic losses caused by chronic exposure.La piel de chinchilla es una de las más exóticas y apreciadas en el mercado internacional. La cría de estos animales es una actividad muy rentable. En Argentina, se han detectado casos de aflatoxicosis aguda en criaderos. Los objetivos de este trabajo fueron: i) estudiar la micobiota y los hongos productores de aflatoxina B1 (AFB1 ) presentes en alimento para chinchillas. ii) analizar la contaminación natural con AFB1 de estos alimentos iii) buscar lesiones histopatológicas en hígados de chinchillas de los criaderos. Se recolectaron muestras de diferentes alimentos (A: chinchilla piel, B: chinchilla madre, C: cubos de alfalfa) en una fábrica y un criadero localizados en la ciudad de Rio Cuarto, en la región central de Argentina. Los hígados de las chinchillas sacrificadas en el criadero fueron analizados macroscópica y microscópicamente. Los recuentos fúngicos totales fueron mayores a 1x104 UFC g-1. Aspergillus, Fusarium y Penicillium fueron los géneros prevalentes, mientras que A. flavus, A. fumigatus, F. verticillioides y F. proliferatum fueron las especies aisladas con mayor frecuencia. 50 % de las cepas de A. flavus aisladas de la fábrica y 69.7 % de las aisladas del criadero produjeron 2.78 a 8.64 µg g-1 y 0.66 a 58.8 µg g-1 de AFB1 , respectivamente. Se detectó AFB1 sólo en las muestras del criadero, con mayor incidencia en el alimento C. Los niveles de toxina variaron entre 1.90 y 97.34 µg kg-1. Los niveles promedios en A y C fueron superiores a 20 µg kg-1. El análisis macroscópico de los hígados reveló apariencia, tamaño y color normal. El análisis microscópico indicó que 63.3 % de los hígados presentaron degeneración lipídica leve a moderada con vacuolización difusa del citoplasma, 9 % presentaron vacuolización lipídica intensa y 27.3 % degeneración hidrópica y vacuolización nuclear en los hepatocitos. El monitoreo periódico de La calidad de los alimentos e ingredientes usados en la alimentación de chinchillas puede evitar intoxicaciones agudas y pérdidas económicas causadas por la exposición crónica a aflatoxinas.Fil: Landa, Maria Florencia. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gonzalez Pereyra, Maria Laura. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Pena, Gabriela Alejandra. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Bagnis, Guillermo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; ArgentinaFil: Cavaglieri, Lilia Reneé. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Rosa, Carlos Alberto da Rocha. Conselho Nacional de Pesquisas Científicas; BrasilFil: Dalcero, Ana Maria. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin